Gold Nanoparticles‐β‐Cyclodextrin‐Chitosan‐Graphene Modified Glassy Carbon Electrode for Ultrasensitive Detection of Dopamine and Uric Acid

The nanocomposite (denoted as GR‐AuNPs‐CD‐CS) of graphene (GR), gold nanoparticles (AuNPs), chitosan (CS) and β‐cyclodextrin (β‐CD) was prepared to modify a glassy carbon electrode. The as‐modified electrode was explored for the ultrasensitive detection of dopamine (DA) and uric acid (UA). The modified electrode demonstrated linearly increased current response in the concentration range of 0.1–120 µm for DA and 0.05–70 µm for UA, with so far the best detection limit for DA and UA. Good stability and repeatability were further demonstrated for the as‐made sensor.     

Gold Nanoparticle Assemblies on Surfaces: Reactivity Tuning through Capping‐Layer and Cross‐Linker Design

The immobilization of metal nanoparticles (NPs) with molecular control over their organization is challenging. Herein, we report the formation of molecularly cross‐linked AuNP assemblies using a layer‐by‐layer approach. We observed four types of assemblies: 1) small aggregates of individual AuNPs, 2) large aggregates of individual AuNPs, 3) networks of fused AuNPs, and 4) gold islands. Interestingly, these assemblies with the different cross‐linkers and capping layers represent different stages in the complete fusion of AuNPs to afford islands of continuous gold. We demonstrate that the stability toward fusion of the nanoparticles of the on‐surface structures can be controlled by the reactivity of the cross‐linkers and the hydrophilicity/hydrophobicity of the nanoparticles.     

Discovery of Chemicals to Either Clear or Indicate Amyloid Aggregates by Targeting Memory‐Impairing Anti‐Parallel Aβ Dimers

Amyloid‐β (Aβ) oligomers are implicated in Alzheimer disease (AD). However, their unstable nature and heterogeneous state disrupts elucidation of their explicit role in AD progression, impeding the development of tools targeting soluble Aβ oligomers. Herein parallel and anti‐parallel variants of Aβ(1–40) dimers were designed and synthesized, and their pathogenic properties in AD models characterized. Anti‐parallel dimers induced cognitive impairments with increased amyloidogenesis and cytotoxicity, and this dimer was then used in a screening platform. Through screening, two FDA‐approved drugs, Oxytetracycline and Sunitinib, were identified to dissociate Aβ oligomers and plaques to monomers in 5XFAD transgenic mice. In addition, fluorescent Astrophloxine was shown to detect aggregated Aβ in brain tissue and cerebrospinal fluid samples of AD mice. This screening platform provides a stable and homogeneous environment for observing Aβ interactions with dimer‐specific molecules.     

Maintenance of Amyloid β Peptide Homeostasis by Artificial Chaperones Based on Mixed‐Shell Polymeric Micelles

The disruption of Aβ homeostasis, which results in the accumulation of neurotoxic amyloids, is the fundamental cause of Alzheimer’s disease (AD). Molecular chaperones play a critical role in controlling undesired protein misfolding and maintaining intricate proteostasis in vivo. Inspired by a natural molecular chaperone, an artificial chaperone consisting of mixed‐shell polymeric micelles (MSPMs) has been devised with tunable surface properties, serving as a suppressor of AD. Taking advantage of biocompatibility, selectivity toward aberrant proteins, and long blood circulation, these MSPM‐based chaperones can maintain Aβ homeostasis by a combination of inhibiting Aβ fibrillation and facilitating Aβ aggregate clearance and simultaneously reducing Aβ‐mediated neurotoxicity. The balance of hydrophilic/hydrophobic moieties on the surface of MSPMs is important for their enhanced therapeutic effect.     

A Cantilever‐based Biosensor for Real‐time Monitoring of Interactions between Amyloid‐β(1–40) and Membranes Comprised of Phosphatidylcholine Lipids with Different Hydrophobic Acyl Chains

Accumulating evidence suggests that interaction between amyloid‐β (Aβ) and cell membrane is crucial to the pathogenesis of Alzheimer's disease (AD), and thus an increasing understanding of the impact of membrane composition on Aβ‐membrane interaction becomes essential for the mechanism elucidation of Aβ‐membrane interaction and the early diagnosis of AD. In this work, electrically neutral phosphatidylcholine (PC) as the most major class of membrane phospholipids, including 1,2‐dipalmitoyl‐sn‐glycero‐3‐phosphocholine (DPPC), 1,2‐distearoyl‐sn‐glycero‐3‐phosphocholine (DSPC), 1‐palmitoyl‐2‐oleoyl‐sn‐glycero‐3‐phosphocholine (POPC), and Aβ(1–40) as the most common amyloid protein were selected as the research subjects, and a developed cantilever‐based biosensor, on which liposomes comprised of PC lipids were immobilized, was applied to characterize in real time the interactions between Aβ(1–40) and membranes comprised of PC lipids with different hydrophobic acyl chains, and to evaluate the effect of cholesterol incorporated in membrane on Aβ‐membrane interaction during the whole process of Aβ(1–40) fibrillization. The results illustrate that the interaction between Aβ(1–40) and PC membrane can be divided into three stages, which are related to the change in molecular states of Aβ. More importantly, it is found that membranes comprised of PC lipids with shorter saturated acyl chains show higher interaction ability with Aβ(1–40), and the incorporation of cholesterol into PC bilayer can remarkably accelerate and strengthen Aβ(1–40)‐membrane interaction. These results confirm that hydrophobicity is the main driving force for the interactions between Aβ(1–40) and PC membranes. In return, the above results enlightened us to apply the current micro‐cantilever immobilized with cholesterol‐containing DPPC liposomes to challenge the detection of low‐concentration Aβ(1–40). This time 50‐nM Aβ(1–40) in aqueous solution has been effectively detected, suggesting that this proposed detection technique would contribute to Aβ detection and early diagnosis of AD in the future.     

Rationally Designed CeNP@MnMoS4 Core‐Shell Nanoparticles for Modulating Multiple Facets of Alzheimer's Disease

Alzheimer's disease (AD) is a complicated multifactorial syndrome. Lessons have been learned through failed clinical trials that targeting multiple key pathways of the AD pathogenesis is necessary to halt the disease progression. Here, we construct core‐shell nanoparticles (CeNP@MnMoS₄) targeting multiple key pathways of the AD pathogenesis, including elimination of toxic metal ions, decrease of oxidative stress, and promotion of neurite outgrowth. The SOD activity and copper removal capacity of CeNP@MnMoS₄‐n (n represents the number of layers of MnMoS₄, n=1–5) was investigated in vitro. We found that CeNP@MnMoS₄‐3 made an excellent balance between SOD activity and copper removal capacity. The effect of CeNP@MnMoS₄‐3 on Cu²⁺‐induced Aβ aggregation was studied by gel electrophoresis, transmission electron microscope (TEM), and atomic force microscopy (AFM). Compared with MnMoS₄ or CeNP alone, a synergistic effect was observed. Moreover, CeNP@MnMoS₄‐3 promoted neurite outgrowth in a dose‐dependent manner. Taken together, the results reported in this work show the potential of new multifunctional core‐shell nanoparticles as AD therapeutics.     

Studies on the cross‐interaction between hIAPP and Aβ25–35 and the aggregation process in binary mixture by electrospray ionization‐ion mobility‐mass spectrometry

A wealth of epidemiological evidence indicates a strong link between type 2 diabetes (T2D) and Alzheimer's disease (AD). The fiber deposition with cross‐β‐sheet structure formed by self‐aggregation and misfolding of amyloidogenic peptides is a common hallmark of both diseases. For the patients with T2D, the fibrils are mainly found in the islets of Langerhans that results from the accumulation of human islet amyloid polypeptide (hIAPP). The major component of aggregates located in the brain of AD patients is amyloid‐β (Aβ). Many biophysical and physiological properties are shared by hIAPP and Aβ, and both peptides show similar cytotoxic mechanisms. Therefore, it is meaningful to investigate the possible cross‐interactions of hIAPP and Aβ in both diseases. In this article, the segment 25–35 of Aβ was selected because Aβ_25–35 was a core region in the process of amyloid formation and showed similar aggregation tendency and toxicity with full‐length Aβ. The electrospray ionization‐ion mobility‐mass spectrometry analysis and thioflavin T fluorescence kinetic analysis combined with transmission electron microscopy were used to explore the effects of the coexistence of Aβ_25–35 and hIAPP on the self‐aggregation of both peptides and whether there was co‐assembly in fibrillation. The results indicated that the aggregation of hIAPP and Aβ_25–35 had two nucleation stages in the binary mixtures. hIAPP and Aβ_25–35 had a high binding affinity and a series of hetero‐oligomers formed in the mixtures of hIAPP and Aβ_25–35 in the early stage. The cross‐reaction between hIAPP monomers and Aβ_25–35 monomers as well as a little of oligomers during primary nucleation stage could accelerate the aggregation of Aβ_25–35. However, owing to the obvious difference in aggregation ability between hIAPP and Aβ_25–35, this cross‐interaction had no significant impact on the self‐assembly of hIAPP. Our study may offer a better understanding for exploring the molecular mechanism of the association between AD and T2D observed in clinical and epidemiological studies and developing therapeutic strategies against amyloid diseases.     

Thermo‐triggered Release of CRISPR‐Cas9 System by Lipid‐Encapsulated Gold Nanoparticles for Tumor Therapy

CRISPR/Cas9 system is a powerful toolbox for gene editing. However, the low delivery efficiency is still a big hurdle impeding its applications. Herein, we report a strategy to deliver Cas9‐sgPlk‐1 plasmids (CP) by a multifunctional vehicle for tumor therapy. We condensed CPs on TAT peptide‐modified Au nanoparticles (AuNPs/CP, ACP) via electrostatic interactions, and coated lipids (DOTAP, DOPE, cholesterol, PEG2000‐DSPE) on the ACP to form lipid‐encapsulated, AuNPs‐condensed CP (LACP). LACP can enter tumor cells and release CP into the cytosol by laser‐triggered thermo‐effects of the AuNPs; the CP can enter nuclei by TAT guidance, enabling effective knock‐outs of target gene (Plk‐1) of tumor (melanoma) and inhibition of the tumor both in vitro and in vivo. This AuNPs‐condensed, lipid‐encapsulated, and laser‐controlled delivery system provides a versatile method for high efficiency CRISPR/Cas9 delivery and targeted gene editing for treatment of a wide spectrum of diseases.     

Label‐free Electrochemical Sensor for Ex‐vivo Monitoring of Alzheimer's Disease Biomarker

The beta‐amyloid (Aβ) peptide was used as an important biomarker for Alzheimer's disease (AD) diagnosis. The development of an accurate, selective, rapid, and highly sensitive technique for detecting of Aβ level is an important issue in biology, and medicine to assess human health risks. Here, gold nanoparticles (Au NPs) with different size were electrochemically deposited onto the indium tin oxide (ITO) substrate in the presence of different molecular weights of surfactants. The modified substrates were used as a high sensitive electrochemical sensor of in‐vitro as well as ex‐vivo monitoring of Aβ based on cyclic voltammetry and square wave voltammetry techniques. Our findings revealed that the modification of ITO electrode with Au NPs could enhance its sensor performance with high sensitivity for low concentration levels of Aβ over a wide linear range with a detection limit of about 20.7 ng/g, which is less than the concentration of insoluble Aβ40 (105.4±40.2 μg/g) in brain of AD induced. In addition, Au NPs/ITO modified electrodes have demonstrated ability to monitor Aβ in the brain extracted samples without any potential interference with other components. Raman spectroscopy has been used to confirm the presence of Aβ in the AD‐induced samples. Thus, it is applicable for analyzing ex‐vivo samples.     

Near-Infrared Aggregation-Induced Emission Luminogens for In Vivo Theranostics of Alzheimer's Disease

Optimized theranostic strategies for Alzheimer's disease (AD) remain almost absent from bench to clinic. Current probes and drugs attempting to prevent β-amyloid (Aβ) fibrosis encounter failures due to the blood–brain barrier (BBB) penetration challenge and blind intervention time window. Herein, we design a near-infrared (NIR) aggregation-induced emission (AIE) probe, DNTPH, via balanced hydrophobicity-hydrophilicity strategy. DNTPH binds selectively to Aβ fibrils with a high signal-to-noise ratio. In vivo imaging revealed its excellent BBB permeability and long-term tracking ability with high-performance AD diagnosis. Remarkably, DNTPH exhibits a strong inhibitory effect on Aβ fibrosis and promotes fibril disassembly, thereby attenuating Aβ-induced neurotoxicity. DNTPH treatment significantly reduced Aβ plaques and rescued learning deficits in AD mice. Thus, DNTPH serves as the first AIE in vivo theranostic agent for real-time NIR imaging of Aβ plaques and AD therapy simultaneously.     

Amphiphilic Janus Gold Nanoparticles Prepared by Interface‐Directed Self‐Assembly: Synthesis and Self‐Assembly

Materials with Janus structures are attractive for wide applications in materials science. Although extensive efforts in the synthesis of Janus particles have been reported, the synthesis of sub‐10 nm Janus nanoparticles is still challenging. Herein, the synthesis of Janus gold nanoparticles (AuNPs) based on interface‐directed self‐assembly is reported. Polystyrene (PS) colloidal particles with AuNPs on the surface were prepared by interface‐directed self‐assembly, and the colloidal particles were used as templates for the synthesis of Janus AuNPs. To prepare colloidal particles, thiol‐terminated polystyrene (PS‐SH) was dissolved in toluene and citrate‐stabilized AuNPs were dispersed in aqueous solution. Upon mixing the two solutions, PS‐SH chains were grafted to the surface of AuNPs and amphiphilic AuNPs were formed at the liquid–liquid interface. PS colloidal particles decorated with AuNPs on the surfaces were prepared by adding the emulsion to excess methanol. On the surface, AuNPs were partially embedded in the colloidal particles. The outer regions of the AuNPs were exposed to the solution and were functionalized through the grafting of atom‐transfer radical polymerization (ATRP) initiator. Poly[2‐(dimethamino)ethyl methacrylate] (PDMAEMA) on AuNPs were prepared by surface‐initiated ATRP. After centrifugation and dissolving the colloidal particles in tetrahydrofuran (THF), Janus AuNPs with PS and PDMAEMA on two hemispheres were obtained. In acidic pH, Janus AuNPs are amphiphilic and are able to emulsify oil droplets in water; in basic pH, the Janus AuNPs are hydrophobic. In mixtures of THF/methanol at a volume ratio of 1:5, the Janus AuNPs self‐assemble into bilayer structures with collapsed PS in the interiors and solvated PDMAEMA at the exteriors of the structures.     

Triterpenoid‐Based Self‐Healing Supramolecular Polymer Hydrogels Formed by Host–Guest Interactions

Pentacyclic triterpenoids, a class of naturally bioactive products having multiple functional groups, unique chiral centers, rigid skeletons, and good biocompatibility, are ideal building blocks for fabricating versatile supramolecular structures. In this research, the natural pentacyclic triterpenoid glycyrrhetinic acid (GA) was used as a guest molecule for β‐cyclodextrin (β‐CD) to form a GA/β‐CD (1:1) inclusion complex. By means of GA and β‐CD pendant groups in N,N′‐dimethylacrylamide copolymers, a supramolecular polymer hydrogel can be physically cross‐linked by host–guest interactions between GA and β‐CD moieties. Moreover, self‐healing of this hydrogel was observed and confirmed by step‐strain rheological measurements, whereby the maximum storage modulus occurred at a [GA]/[β‐CD] molar ratio of 1:1. Additionally, these polymers displayed outstanding biocompatibility. The introduction of a natural pentacyclic triterpenoid into a hydrogel system not only provides a biocompatible guest–host complementary GA/β‐CD pair, but also makes this hydrogel an attractive candidate for tissue engineering.     

Label‐Free,Non‐Derivatization CRET Detection Platform for 6‐Mercaptopurine Based on the Distance‐Dependent Optical Properties of Gold Nanoparticles

A label‐free, non‐derivatization chemiluminescence resonance energy transfer (CRET) detection platform has been developed for the detection of the non‐fluorescent small molecule 6‐mercaptopurine. This CRET process arose from a chemiluminescent (CL) donor–acceptor system in which the reaction of bis(2,4,6‐trichlorophenyl)oxalate (TCPO)–H₂O₂–fluorescein (maximum emission at 521.6 nm) served as the donor and gold nanoparticles (AuNPs, maximum absorption at 520.0 nm) served as the acceptor. This process caused a significant decrease in the CL signal of the TCPO–H₂O₂–fluorescein reaction. The presence of 6‐mercaptopurine induced an aggregation of AuNPs with the assistance of Cu²⁺ ions through cooperative metal–ligand interactions that was accompanied by a distinct change in color and optical properties. The maximum absorption band of the AuNPs was red‐shifted to 721.0 nm and no longer overlapped with the CL spectrum of the reaction; as a result, the CL signal was restored. This CRET system exhibited a wide linear range, from 9.0 nmol L^?1 to 18.0 μmol L^?1, and a low detection limit (0.62 nmol L^?1) for 6‐mercaptopurine. The applicability of the proposed CRET system was evaluated by analysis of 6‐mercaptopurine in spiked human plasma samples.     

Attenuation of the Aggregation and Neurotoxicity of Amyloid‐β Peptides by Catalytic Photooxygenation

Alzheimer’s disease (AD), a progressive severe neurodegenerative disorder, is currently incurable, despite intensive efforts worldwide. Herein, we demonstrate that catalytic oxygenation of amyloid‐β peptides (Aβ) might be an effective approach to treat AD. Aβ1–42 was oxygenated under physiologically‐relevant conditions (pH 7.4, 37 °C) using a riboflavin catalyst and visible light irradiation, with modifications at the Tyr¹⁰, His¹³, His¹⁴, and Met³⁵ residues. The oxygenated Aβ1–42 exhibited considerably lower aggregation potency and neurotoxicity compared with native Aβ. Photooxygenation of Aβ can be performed even in the presence of cells, by using a selective flavin catalyst attached to an Aβ‐binding peptide; the Aβ cytotoxicity was attenuated in this case as well. Furthermore, oxygenated Aβ1–42 inhibited the aggregation and cytotoxicity of native Aβ.     

Tumour‐Targeted Drug Delivery with Mannose‐Functionalized Nanoparticles Self‐Assembled from Amphiphilic β‐Cyclodextrins

Multivalent mannose‐functionalized nanoparticles self‐assembled from amphiphilic β‐cyclodextrins (β‐CDs) facilitate the targeted delivery of anticancer drugs to specific cancer cells. Doxorubicin (DOX)‐loaded nanoparticles equipped with multivalent mannose target units were efficiently taken up via receptor‐mediated endocytosis by MDA‐MB‐231 breast cancer cells that overexpress the mannose receptor. Upon entering the cell, the intracellular pH causes the release of DOX, which triggers apoptosis. Targeting by multivalent mannose significantly improved the capability of DOX‐loaded nanoparticles to inhibit the growth of MDA‐MB‐231 cancer cells with minimal side effects in vivo. This targeted and controlled drug delivery system holds promise as a nanotherapeutic for cancer treatment.     

The Contrasting Recognition Behavior of β‐Cyclodextrin and Its Sulfobutylether Derivative towards 4′,6‐Diamidino‐2‐phenylindole

The noncovalent interactions between 4′, 6‐diamidino‐2‐phenylindole (DAPI) and sulfobutylether β‐cyclodextrin (SBE₇β‐CD) are evaluated by using photochemical measurements and compared with that of native β‐CD. Contrasting recognition behavior and intriguing modulations in the photochemical behavior of DAPI were observed. In particular, a large enhancement in the fluorescence emission and excited‐state lifetime were seen upon binding to SBE₇β‐CD, with the SBE₇β‐CD inclusion complex being approximately 1000 times stronger than that of β‐CD. The ensuing fluorescence “turn on” was demonstrated to be responsive to chemical stimuli, such as metal ions and adamantylanmine (AD). Upon addition of Ca²⁺/AD, nearly quantitative dissociation of the complex was established to regenerate the free dye and result in fluorescence “turn off”. The SO₃^? groups are believed to be critical for the strong and selective binding of the chromophore and the stimuli‐responsive tuning. This is as an important design criterion for the optimization of host–guest properties through supramolecular association, which is relevant for drug‐delivery applications.     

Polymer Cages as Universal Tools for the Precise Bottom‐Up Synthesis of Metal Nanoparticles

A template synthesis allows the preparation of monodisperse nanoparticles with high reproducibility and independent from self‐assembly requirements. Tailor‐made polymer cages were used for the preparation of nanoparticles, which were made of cross‐linked macromolecules with pendant thiol groups. Gold nanoparticles (AuNPs) were prepared in the polymer cages in situ, by using different amounts of cages versus gold. The polymer cages exhibited a certain capacity, below which the AuNPs could be grown with excellent control over the size and shape. Control experiments with a linear diblock copolymer showed a continuous increase in the AuNP size as the gold feed increased. This completely different behavior regarding the AuNP size evolution was attributed to the flexibility of the polymer chain depending on cross‐linking. Moreover, the polymer cages were suitable for the encapsulation of AgNPs, PdNPs, and PtNPs by the in situ method.     

Organoplatinum‐Substituted Polyoxometalate Inhibits β‐amyloid Aggregation for Alzheimer's Therapy

Aggregated β‐amyloid (Aβ) is widely considered as a key factor in triggering progressive loss of neuronal function in Alzheimer's disease (AD), so targeting and inhibiting Aβ aggregation has been broadly recognized as an efficient therapeutic strategy for curing AD. Herein, we designed and prepared an organic platinum‐substituted polyoxometalate, (Me₄N)₃[PW₁₁O₄₀(SiC₃H₆NH₂)₂PtCl₂] (abbreviated as Pt^II‐PW₁₁) for inhibiting Aβ₄₂ aggregation. The mechanism of inhibition on Aβ₄₂ aggregation by Pt^II‐PW₁₁ was attributed to the multiple interactions of Pt^II‐PW₁₁ with Aβ₄₂ including coordination interaction of Pt²⁺ in Pt^II‐PW₁₁ with amino group in Aβ₄₂, electrostatic attraction, hydrogen bonding and van der Waals force. In cell‐based assay, Pt^II‐PW₁₁ displayed remarkable neuroprotective effect for Aβ₄₂ aggregation‐induced cytotoxicity, leading to increase of cell viability from 49 % to 67 % at a dosage of 8 μm . More importantly, the Pt^II‐PW₁₁ greatly reduced Aβ deposition and rescued memory loss in APP/PS1 transgenic AD model mice without noticeable cytotoxicity, demonstrating its potential as drugs for AD treatment.     

Preparation of glycopolymer‐substituted gold nanoparticles and their molecular recognition

Glycopolymer‐substituted gold nanoparticles were prepared via living radical polymerization with a reversible addition‐fragmentation chain transfer (RAFT) reagent. Polyacrylamide derivatives with α‐mannose (α‐Man) and N‐acetyl‐β‐glucosamine (β‐GlcNAc) were synthesized and hydrogenated to obtain thiol‐terminated polymer. The thiol‐terminated glycopolymers were mixed with gold nanoparticles to yield the polymer substituted gold nanoparticles with various diameters, which aggregated on addition of saccharide‐recognition proteins (lectins). The aggregation properties were analyzed using transmission electron microscopy and UV spectra. Molecular recognition was studied with E. coli, which induced aggregation of the nanoparticles at the cell periphery. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 1412–1421, 2009     

Cyclic RGD–Polyethylene Glycol–Polyethylenimine for Intracranial Glioblastoma‐Targeted Gene Delivery

Even though the blood–brain barrier (BBB) is compromised for angiogenesis, therapeutic agents for glioblastoma multiforme (GBM) are particularly inefficient due to the existence of a blood–tumor barrier (BTB), which hampers tumor accumulation and uptake. Integrin α_vβ₃ is overexpressed on glioblastoma U87 cells and neovasculture, thus making its ligands such as the RGD motif target glioblastoma in vitro and in vivo. In the present work, we have designed a modified polyethylene glycol–polyethylenimine (PEG–PEI) gene carrier by conjugating it with a cyclic RGD sequence, c(RGDyK) (cyclic arginine‐glycine‐aspartic acid‐D ‐tyrosine‐lysine). When complexed with plasmid DNA, this gene carrier, termed RGD–PEG–PEI, formed homogenous nanoparticles with a mean diameter of 73 nm. These nanoparticles had a high binding affinity with U87 cells and facilitated targeted gene delivery against intracranial glioblastoma in vivo, thereby leading to a higher gene transfer efficiency compared to the PEG–PEI gene carrier without RGD decoration. This intracranial glioblastoma‐targeted gene carrier also enhanced the therapeutic efficacy of pORF‐hTRAIL, as evidenced by a significantly prolonged survival of intracranial glioblastoma‐bearing nude mice. Considering the contribution of glioblastoma neovasculature to the BBB under angiogenic conditions, our results demonstrated the therapeutic feasibility of treating a brain tumor through mediation of integrin α_vβ₃, as well as the potential of using RGD–PEG–PEI as a targeted gene carrier in the treatment of intracranial glioblastoma.